Collaborating with Team Aachen
Like in all other fields of modern science, collaboration between different work groups is a beneficial way to combine experience and knowledge in order to achieve proper project results. For that reason, iGEM teams from all over the world collaborate to help each other with complex and challenging tasks on their way of accomplishing their specific project aims. In this year, our team joined forces with Aachen in order to improve both of our projects.
Since the primary purpose of our project was to design and create an artificial compartment, orthogonal peroxisomal protein import is one major part of achieving our objective. For that reason, we needed to deactivate natural protein import facilitated by the two receptor proteins Pex5 and Pex7. Initially, we possessed yeast strains which are deficient for Pex5 or Pex7, but we additionally planned to create a yeast cell line which is unable to express both import proteins, resulting in peroxisomes lacking luminal proteins which are called ‘ghosts’. Subsequently, we will integrate our own protein translocation system, allowing us to completely control the protein content of our artificial compartment. We consulted iGEM Aachen which already had lots of experience and knowledge regarding gene knockouts in Saccharomyces cerevisiae. The double knockout was performed successfully by team Aachen, allowing us to get rid of the natural peroxisomal protein import machinery. Conclusively, through this collaboration with team Aachen we were able to improve our final results significantly.
Thanks to the double knockout for the import proteins Pex5 and Pex7 we obtained completely empty ghosts. Thus, verification of the function of our orthogonal peroxisomal protein import machinery was feasible. We therefore accomplished to transfer our artificial Pex5 ‘artico’-receptor including our modified binding pocket region recognizing our modelled PTS1 peptide (PTS1*). When fused to the fluorescent marker protein mTurquoise, our artificial Pex5 facilitates peroxisomal import of the mTurquoise-PTS1* fusion protein indicated by 3D-Sim microscopy.
In return to the help we received from team Aachen we were able to provide them with different plasmids to significantly improve their final project. In the beginning of our collaboration with team Aachen we heard about their problems with building optimized yeast plasmids. For that reason, we introduced them to the yeast toolbox we used which enables the possibility to simplify the process of engineering yeast by creating completely designed plasmids facilitating optimal gene expression and translation for particular purposes in S. cerevisiae. With our golden gate cloning assembly, we designed four different plasmids enabling testing of different promoter strengths for a chloride channel from Arabidopsis thaliana (ATCLCc). We successfully built three yeast optimized plasmids containing the ATCLCc gene with different promotors (Gal1, Sac6, RPL18B) and a fourth plasmid for genome integration containing URA 3’ and 5’ homologies. Correctness of each plasmid was verified by performing restriction digests and sanger sequencing, respectively.
Once we finished the cloning and verification of each plasmid we gave them to team Aachen, allowing them to improve their project results significantly by providing the availability of another vacuolar anion transporter. Thanks to our effort, iGEM Aachen was able to utilize this chloride channel in their yeast based system used for water preparation properly.
Optogenetics offers high spatiotemporal control of well defined events within living cells and therefore qualifies for a broad range of synthetic biological applications. Duesseldorf's last team successfully used optogenetics to induce apoptosis in cancer cells and given that experience, we decided to arrange an optogenetics seminar to share our knowledge with other teams. With that, we hope to reach as many iGEM teams as possible in order to provide essentials in performance of optogenetic experiments. Additionally, we wanted to extend our iGEM teams “toolbox” with one more tool of control.
Our offer found international approval by the teams of Darmstadt, Heidelberg, Paris and Taiwan. Since the teams of Paris and Taiwan could not attend the seminar in person, they joined us via skype.
The seminar started with a marvelous introduction given by Patrick Fischbach. Afterwards, we took advantage of the situation and presented each team’s project, followed by breathtaking discussions facing the difficulties and odds of optogenetic in their project. Introducing the practical handling with optogenetics, Tim Blomeier, who is an advisor of our team, gave a talk about working with optogenetics. With that we imparted all necessary aspects one has to consider when working with optogenetics. Nevertheless nothing is more helpful than practical experience - to attain that, we showed the attendants our laboratory, as well as our hardware and the dark room for optogenetic experiments. At the end of the day we had a nice barbecue. This comfortable situation offers the possibility for lot of interesting discussions regarding optogenetic, our projects and iGEM itself.
Mentoring iGEM Team Stuttgart
The iGEM team of the University of Stuttgart is the first ever at their university. As we are only the second iGEM team of the Heinrich- Heine- University, Duesseldorf, and joined forces with the University of Cologne, which is completely new to iGEM as well, we still remember how hard it can be to meet the iGEM requirements. We found out that they built their team and contacted them to find out how they were getting started on iGEM. After a few friendly mails we decided to schedule a Skype meeting where we talked about many different kinds of topics. We stayed in touch and helped them as much as we could. Furthermore we invited them to the Synbio Day we organized, not only to get another team on board for our mission, but also to finally meet them in person.
As we also knew that they couldn’t afford to come to Boston with their whole team and we had one redundant registration space due to the absence of one of our members, we decided to give this space to Team Stuttgart. We are very happy that iGEM Stuttgart gladly accepted our offer and we cannot wait to see them again at the Giant Jamboree in Boston.
Human Practice collaboration with Valencia UPV
iGEM is not just about working in the lab but also about informing the public and discussing their worries. Therefore we participated in the Human Practice collaboration of Valencia UPV .
They made a worldwide study about the situation regarding transgenics, including various topics like the legislation of transgenic consumption in different countries. Each participating team wrote an essay explaining various aspects of this topic. Different questions were posed such as the social perception of transgenic consumption as well as the viability of growing transgenic plants at home or public spaces. We were glad to participate in this Human Practice collaboration because we think that this is a great way of joining forces to tackle problems like the oftentimes negative social perception of transgenic organisms.
Translation collaboration of Team Franconia
Many teams met at the Germany iGEM meet-up in Dresden. For this meet up iGEM team Dresden organized a Collaboration speeddating. This gave us the opportunity to get in touch with iGEM team Franconia, who were developing a card game. To extend the range of this game, the game was supposed to be translated into many different languages. Our team gladly took part in the effort and translated the game and its instructions to 5 languages: hindu, greek, french, german and portuguese.